Materials and methods for treatment of hepatitis c

ABSTRACT

The present invention relates to materials and methods for treatment of hepatitis C. More closely, the invention relates to human monoclonal antibodies against HCV E1 antigen, to a reagent comprising such antibodies, and to vaccine compositions comprising such antibodies. Futhermore, the invention relates to a method of treating or preventing HCV infection by administration of a vaccine composition comprising the monoclonal antibodies of te invention.

FIELD OF THE INVENTION

[0001] The present invention relates to materials and methods for treatment of hepatitis C. More closely, the invention relates to human monoclonal antibodies against HCV E1 antigen, to a reagent comprising such antibodies, and to vaccine compositions comprising such antibodies for passive immunisation. Furthermore, the invention relates to a method of treating or preventing HCV infection by administration of a vaccine composition comprising the monoclonal antibodies of the invention for passive immunisation.

BACKGROUND OF THE INVENTION

[0002] Human monoclonal antibodies have been assumed as attractive agents for antagonist effects in many medical applications: anti-toxins, anti-receptor molecules, anti-cytokine reagents to reduce or abolish an inflammatory response, etc.

[0003] Hepatitis C virus (HCV) is a major global health problem, with at least 170 million people infected the world over. HCV results in a chronic infection in 75-80% of those initially infected (Houghton 1996). Very likely, immunological factors influence whether the infection will resolve spontaneously or become chronic. The latter will result in a liver inflammation of variable degree, an inflammation that after 10-20 years may result in cirrhosis (20% of chronic cases), and hepatocellular cancer (HCC; approx. 20% of those with cirrhosis) (Houghton 1996). Current pharmaceutical treatment will fail in 60% of the cases (alpha-interferon+Ribavirin). Thus, there is a need for improved therapy.

[0004] HCV was discovered in 1989. Scientific studies have been severely hampered by the fact that there is no robust method to propagate the virus in vitro. Thus, substances cannot be tested for the capacity to block infection by the virus (neutralization assay). Similarly, the only animal model available is chimpanzee, also limiting the number of studies possible (by cost, availability of animals. etc.). As a substitute for a neutralisation assay, the inhibition of binding of one of the two envelope proteins (E2) to target cells has been developed. This is called the NOB assay: neutralisation of binding. Recently, a replicon system where a subgenomic portion of the viral genome replicates inside cells (but is not assembled into viral particles) has been developed, and optimized.

[0005] As mentioned previously, the immune system may have an important role in determining the course of the infection. Both the cellular immune response, as well as the humoral immune system, have been implicated as important for the outcome of the HCV infection. The relative importance of them is still disputed.

[0006] Whether the humoral immune response (i.e. specific B-lymphocytes and antibodies) can interfere with the clinical course of the disease have gained increasing attention over the last years:

[0007] 1. The kinetics of antibodies to the hypervariable region (HVR) of E2 (one of the envelope glyco proteins) may be important: early occurrence of anti-HVR antibodies correlate with resolution of the acute infection.

[0008] 2. Polyclonal anti-HCV immunoglobulin preparations given to liver transplanted patients decreased the occurrence of re-infection by HCV from 94% to 54%.

[0009] 3. Polyclonal anti-HCV given to infected chimpanzees modulated (ameliorated) the course of the infection.

[0010] 4. Individuals lacking immunoglobulin have a tendency to get a more severe and fast progressing disease compared to immunocompetent individuals.

[0011] 5. Anti-HCV antibodies (particularly NOB positive antibodies) correlated with protection in vaccination experiments.

[0012] Accordingly, several groups and companies are currently exploring the possibilities of affecting the course of the infection by administration of anti-HCV antibodies, both to already infected and for prophylaxis (Burioni et al., 1998).

[0013] The present inventors have already cloned human antibodies to conserved regions of one of the two envelope proteins, E2 (Allander et al., 2000). These antibodies have NOB activity, and a patent application for them has been filed, i.e. WO 9740176. They bind to two or possibly three different regions (epitopes) on E2.

[0014] The role of E1 and E2 in the life cycle of the virus is not fully established, nor is the whole process of virus attachment and entry. Still, antibodies to the HVR of E2 can block infection in animals, and so can antibodies to other parts of HCV (most likely E2). Antibodies to E1 elicited by vaccination in chimpanzees correlated with reduced inflammation of the liver (despite constant viral levels in blood); the mechanism for this is unknown (Maertens et al., 2000). There is currently no report within prior art on human monoclonal antibodies to the E1 protein derived from combinatorial libraries.

SUMMARY OF THE INVENTION

[0015] The present inventors have been able to generate human antibodies to the E1 protein. This was much more complicated than isolating antibodies against the E2 protein, as there seem to be an immuno-dominance in most infected individuals to generate anti-E2 antibodies rather than anti-E1 inmunoglobulins. Initially, the inventors worked with a recombinant protein resembling the complexed E1/E2, the assumed native complex presented on the viral surface. Only anti-E2 antibodies could be isolated in those experiments. To solve the problem of generating anti-E1 antibodies, E1 was first cloned and expressed separately on the surface of eukaryotic cells. Subsequently, such E1 displaying cells were used for selection of anti-E1 binding clones from an antibody library displayed on filamentous phage, in order to avoid the immunodominant anti-E2 clones present in the phage antibody library.

[0016] In a first aspect, the invention provides a recombinant human monoclonal antibody, or antigen binding fragments thereof, that exhibits immunological binding affinity for a hepatitis C virus (HCV) E1 antigen, wherein said monoclonal antibody comprises an amino acid sequence homologous to the binding portion of a human antibody Fab molecule obtained from a combinatorial antibody library.

[0017] The monoclonal antibody according to the invention reacts with complexed HCV E1/E2 antigen. The monoclonal antibody according to the invention preferably reacts with genotypically different isolates of HCV virus. Furthermore, the monoclonal antibody according to the invention may be improved by mutation and new selection, i.e. so called affinity maturation in vitro, to increase the binding strength of the antibodies to E1.

[0018] The Fab molecule of the monoclonal antibody of the invention comprises the VH and VL domains respectively, of the heavy and light chains of the Fab molecule according to Seq. ID No. 1-56 of the enclosed Sequence Listning.

[0019] The monoclonal antibody according to the invention may be fused with a further substance for diagnostic or therapeutic purposes, for example a toxin, an antibody allowing targeting, e.g. against defense cells, a protein conferring modulated metabolism of the anti-E1 antibody, a marker for diagnosis, immunohistochemistry, imaging etc.

[0020] In a second aspect, the invention relates to a detecting immunological reagent comprising the monoclonal antibody according to the invention. For example, the reagent may be used in quantitative assays for detection and analysis of replication and assembly in the life cycle of the virus.

[0021] In a third aspect, the invention relates to an immunological assay comprising the above reagent. The assay may be a modified NOB assay as mentioned above. Alternatively, the immunoassay may be a qualitative assay to measure conformation of recombinant E1 in connection with the production of therapeutic and diagnostic agents.

[0022] In a fourth aspect, the invention relates to a drug or vaccine composition against HCV infection, comprising the monoclonal antibody according to the invention or any combination of antibodies, or antibody binding fragments, of the invention. The vaccine compositon is formulated with pharmaceutically acceptable vehicles in a conventional manner and is intended for passive immunisation.

[0023] According to the present invention it is possible to produce human antibodies to parts of E2 (in particular the HVR region), and to the envelope protein, E1, in order to compose a “cocktail” of 3-6 or more human monoclonal antibodies. Such a mixture of antibodies to different proteins and protein regions of the virus has a much larger probability of severely affecting such a variable virus as HCV.

[0024] In a fifth aspect, the invention provides a method of treating a subject against HCV infection in a therapeutic or prophylactic purpose, comprising administration of the vaccine composition according to the invention to subjects in need thereof.

DETAILED DESCRIPTION OF THE INVENTION Materials and Methods

[0025] Antibodies and Antisera

[0026] Monoclonal mouse anti-human-myc from Invitrogen was used in 1:1.50-1:75 dilution (9-19 μg/ml). Ascites produced mouse anti-E1 monoclonal antibody was diluted 1:250. A FITC conjugated rabbit-anti-mouse immunoglobulin from Dako was used as secondary antibody in 1:15-1:10 dilution (67-100 μg/ml).

[0027] Anti-HCV Phage Library Construction

[0028] The library was derived from a bone marrow donation from a patient infected with HCV genotype 1a. Construction of the library was performed as described in Allander et al., 2000. In brief, lymphocytes were isolated from bone marrow using Ficoll-Paque (Pharmacia), and total RNA was extracted by the acid guanidium thiocyanate-phenol method. First strand cDNA synthesis was performed using an oligo-dT primer and the “First strand cDNA synthesis” kit (Amersham Pharmacia). The cDNA was subsequently used as template for for PCR amplification of γ1 Fd and κ light chains (Kang et al., 1991), and ligated into the phagemid vector pComb3H (Barbas and Wagner, 1995).

[0029] Construction of E1 Expression Vector

[0030] The signal sequence and echtodomain of E1 (aa 174 to 359) was inserted into the vector pDisplay (Invitrogen). The pDisplay vector has a mouse Igκ signal sequence upstream of its cloning cassette and a PDGF membrane anchor sequence downstream of the cassette for cell-surface expression. In addition, the vector contains two tag sequences so that the expressed protein will have a N-terminal Hemagglutinin A epitope tag, and a C-terminal myc epitope tag before the membrane anchor. The signal sequence and echtodomain fragment of E1 was PCR amplified from a full-length HCV clone (pcv H77c, genotype 1a), which was a generous gift from Dr J Bucht, NIH, USA (Yanagi et al., 1997). Since the hemagglutinin A tag would be localized N-terminal of the E1 fragment and possibly disturb recognition by anti-E1 or anti-myc antibodies, the signal sequence and the Hemagglutinin tag were deleted from the vector using Eco RI and Pst I (Life Technologies). The E1 fragment was subsequently inserted with the 3′ end in the cloning cassette and the 5′ end replacing the removed sequences. The PCR was performed as follows: first 94° C. 5 min; then 94° C. 1 min, 52° C. 0.5 min, and 72° C. 10 min for 35 cycles; and finally 72° C. 10 min. Primers (Symbion, Denmark) were designed to contain specific restriction enzyme sites for directional cloning and an ATG start codon in the sense primers: sense primer (H77C-S1b) 5′-GG AAC CTT CCT GAA TTC GGC TTG GGG ATG TTC TCT ATC-3′ (restriction site for Eco RI underlined), antisense primer (H77C-AS1) 5′-CAT GGA GAA ATA CGC CTG CAG CGC CAG-3′ (restriction site for Pst I underlined). The PCR fragment was cleaved with the mentioned restriction endonucleases, and gel purified on 1% agarose. The band of correct size (approx. 600 bp) was cut out from the gel, and DNA was eluted using Concert DNA purification kit (Life Technologies). The pDisplay vector (Invitrogen) was cut using the same restriction enzymes and likewise gel purified. The fragment was ligated to the vector in 1:1, 1:3 and 1:6 ratios at +4° C. o.n. using T4 DNA ligase (Life Technologies). The 1:1 ligation product was linearized, self-religated and used to transform E- coli (XL-1 Blue, Stratagene) by electroporation. Single ampicillin resistant clones were picked and cultured, and DNA was extracted using Wizards Plus Midipreps DNA purification system (Promega). To distinguish which clones contained correct insert, DNA from single clones was analyzed both by PCR and by restriction enzyme digestion.

[0031] Detection of E1 Expression on Eukaryotic Cells

[0032] Expression of the E1-myc construct was tested in CHO and HeLa cells. Cells were grown in 6-well plates to near confluence in RMPI 1640 (CHO cells) or DMEM (HeLa cells) supplemented with 10% fetal bovine serum (FBS) and 100 U /ml penicillin, streptomycin 100 μg/ml (Life Technologies). Transfection was performed using FuGENE 6 (Boehringer-Mannheim) with a FuGENE 6 to DNA ratio of 3-6 μl to 1-2 μg per well. Geneticin (Life Technologies) was added to the culture media, in a final concentration of 200 μg/ml, 48 h after transfection. Analysis with immunofluorescence microscope (Leitz DMRBE, Leica) and flowcytometry (FacSort, Becton Dickinson) were carried out on solubilized cells. Cells were loosened using a rubber policeman and washed twice in wash buffer Dulbeccos' PBS, 0.5% FBS, 0.1% sodium azide (Life Technologies, USA). Cells were centrifuged and resuspended in a small volume of primary antibody (anti-myc or anti-E1) and incubated at RT for 60 min. After two to three washes in wash buffer, cells were incubated in the dark at RT for 30 min in secondary rabbit-anti-mouse Ig-FITC. Finally cells were briefly incubated in Hoerst 1:1000, washed 3 times and mounted on glass slides in 5-10 μl Vectashield(Vector). Cells to be analysed with flowcytometry were redisolved in 250 μl washbuffer.

[0033] Selection of Anti-E1 Clones

[0034] Anti-E1 clones were selected from three different sets of selections. For the first two sets, HeLa cells were grown to semi confluence in T75 culture flasks. In the first set of selections, cells were transfected with 8-9 μg E1 DNA (47-53 μl Fugene6). In the second set, HeLa cells were transfected twice with the E1 DNA to further increase the surface expression prior to selection. Cells were first transfected (5.5 μg DNA) in a T25 culture flask for two days, grown under Geneticin selection pressure for 5 days, and then moved and transfected in a T75 flask (16 μg DNA). In the third set, CHO cells were used because of their higher surface expression even after the first transfection. Cells were transfected with 16-30 μg DNA in T75 flasks. One to four days after transfection cells were harvested using a rubber policeman, and suspended in DPBS—4% nonfat milk—0.02% sodium azide. To deplete the phage library of non-specific binders, the phages were incubated with non-transfected cells for one hour on an orbit shaker 100 rpm at RT or incubated overnight at +4° C. and then on a turning-wheel (7 rpm) for one hour at RT. Cells were removed by centrifugation, and the depleted phages were incubated with transfected cells for 1.5-2 h on a turning-wheel at RT. Cells were then washed three times in wash-buffer (Dulbeccos PBS, 0.5% FBS, 0.02% sodium azide). Cells were resuspended in 100 μl anti-myc 1:75 dilution and incubated at RT for 2 h. After two washes the cells were resuspended in FITC-anti-mouse—IgG 1:15 dilution and incubated in the dark for 1 h. The cells were washed twice before resuspended (10⁶ cells/ml) in wash-buffer. Sorting was performed in a FACSVantage SE (Becton Dickinson). Phages were eluted from the sorted cells by adding 200 μl 0.1 M HCl-glycine pH 2.2. Eluted phages were used to infect freshly cultured XL-1 Blue. The infected bacteria were then plated out on LA-amp plates. The next day, colonies were harvested in SB and grown to a 50-100 ml culture. Phages were induced and harvested as described (Barbas et al., 1991), and used for a next round of selection.

[0035] Fab Expression and Initial Testing

[0036] Colonies were picked, propagated and analyzed as single clones. Fab production was induced and a periplasmic fraction was prepared by freeze thawing (Allander et al 2000). Fab production was determined in an ELISA using anti-Fd (The Binding Site, UK) and AP conjugated anti-Fab (PIERCE, USA). Specificity for the antigen was initially tested in an ELISA against an recombinant E1/E2 protein, or against recombinant E1 protein. The recombinant E1/E2 heterodimer protein (genotype 1a) expressed in CHO cells was generously provided by Dr M. Houghton, Chiron Corp. and has been described elsewhere (Spaete et al., 1992). The soluble E1 protein was expressed and secreted into the medium from COS cells, and was generously provided by Dr. A. Patel, MRC Virology Unit, Glasgow, U.K. The ELISA assays with these antigens were performed as described below.

[0037] ELISA Assay for Determination of Specific Binding of the Antibodies to HCV Proteins

[0038] ELISA for E1 reactivity: GNA lectin (Sigma) was diluted to 2.5 μg/ml in PBS and coated to microtiter wells (Costar 3690) over night at room temperature. The wells were washed once in PBS with 0.05% Tween 20 (PBS-T) and the wells were blocked with 4% non-fat dry milk in PBS for 2 hours at room temperature. A:fter discarding the blocking solution, 50 μl of recombinant E1 (approx. concentration 50 μg/ml) was added and incubated for two hours at room temperature. After the E1 solution had been discarded, the antibody preparations were added and bound antibodies detected as described below.

[0039] Recombinant E1/E2 heterodimer (genotype 1a) was diluted to 1 μg/ml in PBS, and coated to microtitre wells overnight at +4° C. Unbound antigen was discarded, and the wells were blocked with 5% non-fat dry milk in PBS for 60 minutes at room temperature. Blocking solution was discarded, and antibody solutions to be tested added in 1:3-1:24 dilutions (diluent: PBS with 0.05% Tween 20). The plates were incubated at 37° C. temperature for 2 hours, washed four times with PBS-T, and alkaline phosphatase (AP) coupled-goat anti-human F(ab′)₂ (Pierce, Rocherford, Ill.) antibodies in a 1:1000 dilution were added. After 60 minutes at 37° and subsequent washes, substrate solution (p-nitrophenyl phosphate, Sigma, St. Louis, Mo.) was added and absorbency measured at 405 nm.

[0040] For control purposes, recombinant E2 (genotype 1a), or BSA (Sigma) coated at 1 μg/ml were used in corresponding ELISAs to control for unspecific reactivity.

[0041] Transfer of Fab Clones into IgG Format

[0042] The Fd and the light chain gene segments were transferred from the phagernide vector pComb3H to the eukaryotic vector pcIgG1 as previously reported (Samuelsson et al., 1996). Plasmid DNA was transfected into CHO cells using Lipofectamine Plus (Life Technologies) according to the manufacturer's instructions. Medium containing secreted IgG was harvested every second day and frozen until analyzed. The ELISA to determine IgG concentration used a rabbit anti-human IgG and an AP conjugated rabbit anti-human IgG, and a purified human IgG standard as reference (Dako) (Samuelsson et al., 1996).

RESULTS

[0043] Library Size

[0044] Ligation of γ1 Fd genes into the pComb3H vector gave a library of 7.8×10⁶ cfu/μg, and ligation of κ light chain genes into the pComb3H gave a library of 1.6×10⁷ cfu/μg. The resulting combinatorial library comprised 3.7×10⁷ members.

[0045] Construction of E1 Expression Vector

[0046] The first ligation of pDisplay vector and E1 fragment resulted in similar number of clones independent of ligation ratios. Plasmid DNA extracted from cultures of the 1:1 ligation was linearized and separated on a gel. Two bands of approximately 5.5 and 6.0 appeared on the gel. Since the expected correct sized band was 5.8 kb, both bands were purified and religated separately. Single clones were grown and insertion of the fragment was demonstrated by PCR using the same sense and antisense primers as in the cloning step; 9 clones out of 20 showed the correct fragment length. Subsequent restriction enzyme digestion showed that the correct sized fragment also could be cleaved from all 9 clones. Nucleic acid sequencing of the clones showed that they all differed from the original E1 sequence by a few nucleotides or more.

[0047] Assessment of E1 Expression on Eucaryotic Cells

[0048] Initially HeLa cells were preferred. To determine optimal expression of the myc-tag, expression was investigated by fluorescence microscopy and flow cytometry on day 1, 2, 4, and 7 after transfection. This time study indicated that immunofluorescence detection of the myc tag was optimal 4 days after transfection. For the second set of selection surface expression was increased slightly by transfecting the HeLa cells twice. For the third set CHO cells were chosen since they appear to be more tolerant to transfection as well as show clear E1 expression already after one day of transfection.

[0049] Selection of E1 Specific Clones

[0050] In the first round of each selection series, approximately 10¹¹ cfiu of phage library was depleted against 10⁶-10⁷ non-transfected cells. Unbound, depleted phages were subsequently incubated with 3-8×10⁶ transfected, E1 expressing cells. After sorting for myc positive cells, phages were eluted from the sorted cells and re-propagated in fresh XL1 blue. 2-8×10¹⁰ cfu of re-propagated phages were used in the next selection round.

[0051] In the first series, two rounds of selection were performed. After the second round, only 108 colonies were formed, 98 of which were tested in ELISAs for Fab expression and binding to recombinant E1/E2 antigen. Sixteen clones expressed Fab, of which nine were positive for binding to E1/E2. After nucleic acid sequencing, two clones were judged not to be proper Fab fragments. Two clones (clones 13 and 98) from this selection series were further characterised and included in the present collection of antibodies (Table 1, Seq ID No. 1-4).

[0052] In the second series, six selection rounds were performed. After the fourth round, 42 single clones were picked and assayed for Fab expression. Twentytwo clones produced Fab, while only two were positive when tested for E1 reactivity. Subsequent sequence analysis revealed that the clones were identical (clone 4:6; Seq ID No. 13-14). An additional clone, isolated from the sixth panning round in this series, was also characterised (clone 6a:5; Table 1 and Seq ID No. 15-16).

[0053] In the third series, two rounds of selection were made and the second round was repeated once. 45-75% of propagated clones expressed Fab. The majority of our E1 specific Fab clones were isolated from this selection series (clones with prefix 1:, 2a: and 2b:).

[0054] Reactivity to HCV Antigens

[0055] The reactivity of the different Fab clones to the HCV proteins E1, E1/E2 in complex, free E2 or BSA was determined by ELISA. All clones showed a significantly higher reactivity to E1 and/or E1/E2 than against E2 or BSA (negative control antigens) (Table 1). From these data, it seems that some clones may be particularly efficient binders: clones 1.4, 1:8, 2a: 13, 2a:23, 2a:30, 2b:5 and 4:6 TABLE 1 Reactivity to HCV antigens and BSA by the Fab proteins (crude periplasmic preparations) as measured by ELISA (OD_(405 nm) values given). For technical reasons, E2 was in some experiments replaced with BSA as negative control antigen (values marked with (B)). n d = not determined μg Fab/ml Fab clone E1 E1/E2 E2 or BSA (B) (approximative) 13 0.20 0.16 0.03 (B) >3 98 0.20 0.16 0.10 (B) 5 1:4 0.78 0.67 0.27 (B) 0.01 1:8 0.75 0.53 0.09 (B) 0.01 1:9 0.96 0.67 0.16 (B) 0.05  1:10 0.20 0.12 0.01 (B) 0.01 4:6 0.97 0.20 0.08 0.24 6a:5  0.15 0.09 0.03 0.33 2a-2  1.00 0.32 0.18 0.64 2a-4  1.18 0.42 0.33 1.0 2a-5  0.89 0.08 0.07 0.44 2a-13 1.05 0.22 0.14 0.36 2a-14 0.36 0.09 0.07 0.72 2a-23 1.47 0.23 0.19 0.50 2a-25 1.46 0.33 0.24 0.11 2a-30 1.11 0.52 0.26 0.96 2a-32 0.61 0.09 0.07 >1 2a-33 0.65 0.14 0.14 >1 2a-37 0.99 0.48 0.29 (B) 0.08 2a-40 1.21 0.17 0.17 1.0 2b-1  0.35 0.12 0.08 0.94 2b-3  1.18 0.19 0.12 1.0 2b-4  0.11 0.09 0.03 (B) 0.68 2b-5  2.21 1.40 0.71 >1.0 2b-7  0.47 0.32 0.18 0.54 2b-9  0.33 0.19 0.15 0.19 2b-10 0.48 0.11 0.13 >1.0 2b-17 0.24 n d 0.05 0.16 mouse mcl anti-E1 >3.00 >3.00 0.20 rabbit anti-E2 n d 0.15 0.72 PBS 0.11 0.10 0.08 non-specific Fab 0.20 0.11 0.14 (B)

[0056] References

[0057] Allander T, Drakenberg K, Beyene A, Rosa D, Abrignani S, Houghton M, Widell A, Grillner L, Persson MAA. Recombinant human monoclonal antibodies against different conformational epitopes of the E2 envelope glycoprotein of hepatitis C virus that inhibit its interactions with CD81. J Gen Virol 2000; 81: 2451-2459.

[0058] Barbas III C F, Kang A S, Lerner R A, Benkovic S J. Assembly of combinatorial antibody libraries on phage surfaces: the gene III site. Proc Natl Acad Sci U S A 1991; 88: 7978-82.

[0059] Barbas III C F, Wagner J. Synthetic human antibodies: selecting and evolving finctional proteins. Methods 1995; 8: 94-103.

[0060] Burioni R, Plaisant P, Manzin A, Rosa D, Delli Carri V, Bugli F, Solforosi L, Abrignani S, Varaldo P E, Fadda G, Clementi M. Dissection of human humoral immune response against hepatitis C virus E2 glycoprotein by repertoire cloning and generation of recombinant Fab fragments. Hepatology 1998; 28: 810-814.

[0061] Houghton M. Hepatitis C Virus. In Fields Virology, (eds B N Fields, D M Knipe, P M Howley) Lippincott-Raven Publishers, Philadelphia, 1996 pp1035-1058.

[0062] Kang A S, Burton D R, Lemer R A. Combinatorial inimunoglobulin libraries in phage λ. Methods: Comp. Methods in Enzymol. 1991;2: 111-8.

[0063] Maertens G, Priem S, Ducatteeuw A, Verschoorl E, Verstrepen B, Roskams T, Desmet V, Fuller S, Van Hoek K, Vandeponseele P, Bosman F, Buyse M A, van Doom L J, Heeney J, Kos A, Depla E. Improvement of chronic active hepatitis C in chronically infected chimpanzees after therapeutic vaccination with the HCV E1 protein. Acta Gastroenterologica Belgica. 2000; 63: 203.

[0064] Samuelsson A, Yari F, Hinkula J, Ersoy O, Norrby E, Persson M A A. Human antibodies from phage libraries: neutralizing activity against human immunodeficiency virus type 1 equally improved after expression as Fab and IgG in mammalian cells. Eur J Immunol 1996; 26: 3029-34.

[0065] Spaete R R, Alexander D, Rugroden M E, Choo Q L, Berger K, Crawford K, Kuo C, Leng S, Lee C, Ralston R, and others. Characterization of the hepatitis C virus E2/NS 1 gene product expressed mammalian cells. Virology 1992; 188: 819-830.

[0066] Yanagi M, Purcell R H, Emerson S U, Bukh J. Transcripts from a single full-length cDNA clone of hepatitis C virus are infectious when directly transfected into the liver of a chimpanzee. Proc Natl Acad Sci USA 1997; 94:8738-8743.

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Ala Ser Val Gly Asp 1 5 10 15 Arg Val Thr Ile Thr Cys Gln Ala Ser Pro Asp Ile Ser Asn Tyr Leu 20 25 30 Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Asn Leu Leu Ile Tyr 35 40 45 Gly Ala Ser Ser Leu Gln Arg Gly Val Pro Ser Arg Leu Ser Gly Ser 50 55 60 Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Asp Ser Leu Gln Pro Glu 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr Gly Ser Pro His Thr 85 90 95 Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 5 125 PRT Homo sapiens PEPTIDE (1)..(125) Clone 14 VH 5 Ala Glu Val Gln Leu Leu Glu Ser Gly Pro Gly Leu Val Lys Pro Ser 1 5 10 15 Glu Thr Leu Ser Leu Thr Cys Thr Val Ser Asn Gly Ser Met Ser Asn 20 25 30 Tyr Cys Trp Ser Trp Ile Arg Gln Ser Pro Gly Lys Gly Leu Glu Trp 35 40 45 Ile Gly Tyr Ile Tyr Tyr Ser Gly Ser Thr Ser Tyr Asn Pro Ser Leu 50 55 60 Arg Ser Arg Val Ala Leu Leu Val Asp Thr Ser Lys Asn Gln Phe Ser 65 70 75 80 Leu Lys Leu Thr Ser Val Thr Thr Ala Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Asp Leu Ser Ala Arg Gly Gly Thr Arg Asn Arg Asp Ala Leu 100 105 110 Asp Val Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ala 115 120 125 6 107 PRT Homo sapiens PEPTIDE (1)..(107) Clone 14 VK 6 Ala Glu Leu Thr Gln Ser Pro Ala Thr Leu Ser Val Ser Pro Gly Glu 1 5 10 15 Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Asn Ser Asn Leu 20 25 30 Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Ser Leu Leu Ile Tyr 35 40 45 Gly Ala Ser Thr Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Glu Leu Thr Leu Thr Ile Ser Ser Leu Gln Ser Glu 65 70 75 80 Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Lys Asn Trp Pro Pro Trp 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 7 127 PRT Homo sapiens PEPTIDE (1)..(127) Clone 18 VH 7 Ala Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Glu Pro Gly 1 5 10 15 Arg Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Asn Phe Gly Asp 20 25 30 Tyr Ala Ile His Trp Val Arg Gln Gly Pro Gly Lys Gly Leu Glu Trp 35 40 45 Val Ala Gly Ile Ser Trp Asn Ser Gly Ser Ile Gly Tyr Ala Asp Ser 50 55 60 Val Lys Gly Arg Phe Ile Ile Ser Arg Asp Asn Ala Lys Lys Thr Leu 65 70 75 80 Phe Leu Gln Met Asn Thr Leu Arg Thr Glu Asp Thr Ala Leu Tyr Tyr 85 90 95 Cys Val Lys Glu Thr Gly Ala Gln Gly Val Ala Gly Ser Gly Ala Tyr 100 105 110 Tyr Phe His Asn Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 125 8 106 PRT Homo sapiens PEPTIDE (1)..(106) Clone 18 VK 8 Ala Glu Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Ile Gly Asp 1 5 10 15 Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Asn Tyr Val 20 25 30 Ala Trp Tyr Gln Gln Arg Pro Gly Lys Val Pro Lys Leu Leu Ile Phe 35 40 45 Gly Ala Ser Ala Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Thr Ser Leu Gln Pro Glu 65 70 75 80 Asp Val Ala Thr Tyr Tyr Cys Gln Lys Tyr Arg Ser Ala Pro Leu Thr 85 90 95 Phe Gly Pro Gly Thr Arg Val Asp Leu Lys 100 105 9 121 PRT Homo sapiens PEPTIDE (1)..(121) Clone 19 VH 9 Ala Glu Val Gln Leu Leu Glu Ser Gly Pro Gly Leu Val Lys Pro Ser 1 5 10 15 Glu Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Asn Ser Ile Ser Ser 20 25 30 Gly Tyr Tyr Trp Val Trp Phe Arg Gln Ser Pro Gly Lys Gly Leu Glu 35 40 45 Trp Ile Ala Ser Ile Tyr His Ser Gly Ser Thr Tyr Tyr Asn Pro Ser 50 55 60 Leu Arg Ser Arg Val Ser Ile Ser Val Asp Thr Ser Lys Lys Gln Phe 65 70 75 80 Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr 85 90 95 Cys Ala Arg Asp Ser Ala Lys Thr Thr Arg Tyr Phe Val His Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 10 109 PRT Homo sapiens PEPTIDE (1)..(109) Clone 19 VK 10 Ala Glu Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly Glu 1 5 10 15 Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Ser Tyr 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Ala Gly Gln Ala Pro Arg Leu Leu Ile 35 40 45 Tyr Gly Ala Ser Arg Arg Ala Thr Gly Ile Pro Val Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Ala Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Ser 65 70 75 80 Glu Asp Ser Ala Val Tyr Tyr Cys Gln His Tyr His Asn Trp Pro Ala 85 90 95 Met Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105 11 132 PRT Homo sapiens PEPTIDE (1)..(132) Clone 110 VH 11 Ala Glu Val Gln Leu Leu Glu Ser Gly Ala Glu Val Lys Lys Pro Gly 1 5 10 15 Glu Ser Leu Arg Ile Ser Cys Arg Gly Ser Gly Tyr Ser Phe Pro Asn 20 25 30 Tyr Trp Val Ser Trp Val Arg Gln Met Pro Gly Lys Gly Leu Glu Trp 35 40 45 Met Gly Lys Ile Asp Pro Ser Asp Ser Glu Thr Asn Tyr Ser Pro Ser 50 55 60 Phe Gln Gly His Val Thr Ile Ser Ala Asp Lys Ser Leu Ser Ile Ala 65 70 75 80 Tyr Leu Gln Trp Asn Ser Leu Lys Ala Ser Asp Thr Ala Met Tyr Tyr 85 90 95 Cys Ala Arg His Lys Arg Gly Ala Pro Thr Tyr Lys Asp Ile Leu Thr 100 105 110 Gly Tyr Tyr Val Asp Gly Met Asp Val Trp Gly Gln Gly Thr Thr Val 115 120 125 Thr Val Ser Ser 130 12 112 PRT Homo sapiens PEPTIDE (1)..(112) Clone 110 VK 12 Ala Glu Leu Thr Gln Ser Pro Asp Ser Leu Ala Met Ser Leu Gly Glu 1 5 10 15 Arg Ala Ser Ile Asn Cys Lys Ser Ser Arg Ser Leu Leu Tyr Ser Ser 20 25 30 Asn Asn Lys Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Ser Gly His Pro 35 40 45 Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val Pro 50 55 60 Asp Arg Phe Asn Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile 65 70 75 80 Ser Ser Leu Gln Ala Glu Asp Val Ala Val Tyr Phe Cys Gln Gln Tyr 85 90 95 Tyr Ser Ser Pro Leu Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100 105 110 13 125 PRT Homo sapiens PEPTIDE (1)..(125) Clone 46 VH 13 Ala Glu Val Gln Leu Leu Glu Gln Ser Gly Ala Glu Val Lys Lys Pro 1 5 10 15 Gly Ala Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr 20 25 30 Lys Tyr Tyr Leu His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu 35 40 45 Trp Met Gly Phe Ile Asn Pro Ser Gly Gly Ser Thr Ser Ser Ala Gln 50 55 60 Lys Phe Gln Gly Arg Ile Ser Met Thr Arg Asp Thr Ser Thr Thr Thr 65 70 75 80 Val Tyr Met Glu Val Asn Ser Val Thr Ser Glu Asp Thr Ala Val Tyr 85 90 95 Tyr Cys Ala Arg Val Gly Arg Leu Gly Val Gly Ala Thr Gly Ala Phe 100 105 110 Asp Leu Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser 115 120 125 14 107 PRT Homo sapiens PEPTIDE (1)..(107) Clone 46 VK 14 Ala Glu Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp 1 5 10 15 Arg Val Thr Ile Thr Cys Gln Ala Ser Gln Asp Ile Asn Tyr Ser Leu 20 25 30 Asn Trp Tyr Gln Gln Lys Pro Gly Lys Pro Pro Lys Leu Leu Ile Tyr 35 40 45 Tyr Ala Ser His Ser Asp Thr Gly Val Pro Ser Arg Phe Ser Gly Gly 50 55 60 Gly Ser Gly Thr Arg Phe Thr Leu Thr Ile Tyr Ser Leu Gln Pro Glu 65 70 75 80 Asp Ile Ala Thr Tyr Tyr Cys Gln His Phe Asp His Val Pro Arg Tyr 85 90 95 Thr Phe Gly Pro Gly Thr Lys Val Asp Leu Lys 100 105 15 128 PRT Homo sapiens PEPTIDE (1)..(128) Clone 6a5 VH 15 Ala Glu Val Gln Leu Leu Glu Ser Gly Pro Gly Leu Val Lys Pro Ser 1 5 10 15 Gln Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Asn Val Ser Arg 20 25 30 Lys Ser Ala Ala Trp Asn Trp Ile Arg Gln Ser Pro Ser Arg Gly Leu 35 40 45 Glu Trp Leu Gly Arg Thr Tyr His Met Ser Lys Trp Tyr Ser Val Tyr 50 55 60 Ala Thr Ser Leu Lys Ser Arg Ile Asn Ile Asn Val Asp Thr Ser Arg 65 70 75 80 Asn Gln Phe Ala Leu Gln Leu Arg Ser Val Thr Pro Glu Asp Thr Ala 85 90 95 Val Tyr Tyr Cys Ala Arg Glu Gly Pro Glu Trp Ala Val Gly Gly Thr 100 105 110 Tyr Gly Met Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser 115 120 125 16 106 PRT Homo sapiens PEPTIDE (1)..(106) Clone 6a5 VK 16 Ala Glu Leu Thr Gln Ser Pro Asp Thr Leu Ser Leu Ser Pro Gly Glu 1 5 10 15 Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Asn Asn Asn Tyr 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile 35 40 45 Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser Gly 50 55 60 Ser Gly Ser Glu Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu Pro 65 70 75 80 Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Asp Ser Ser Arg Thr 85 90 95 Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 17 130 PRT Homo sapiens PEPTIDE (1)..(130) Clone 2a2 VH 17 Ala Glu Val Gln Leu Leu Glu Gln Ser Gly Ala Glu Val Lys Lys Pro 1 5 10 15 Gly Ala Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr 20 25 30 Gly Tyr Tyr Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu 35 40 45 Trp Met Gly Trp Ile Asn Pro Glu Ser Gly Ala Thr Asn Tyr Ala Gln 50 55 60 Asn Phe Gln Gly Arg Val Thr Met Thr Thr Asp Thr Ser Met Arg Thr 65 70 75 80 Ala Tyr Ile Glu Val Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr 85 90 95 Phe Cys Ala Arg Gly Gly Ala Phe Cys Thr Gly Gly Thr Cys Tyr Phe 100 105 110 Ala Ile Tyr Gly Met Asp Val Trp Gly Gln Gly Thr Ala Val Ile Val 115 120 125 Ser Ser 130 18 110 PRT Homo sapiens PEPTIDE (1)..(110) Clone 2a2 VK 18 Ala Glu Leu Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly Glu 1 5 10 15 Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Leu His Thr Asn 20 25 30 Gly Tyr Asn Tyr Leu Asp Trp Tyr Leu Gln Lys Pro Gly Gln Ser Pro 35 40 45 Gln Leu Leu Ile Tyr Leu Gly Ser Asn Arg Ala Ser Gly Val Pro Asp 50 55 60 Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile Ser 65 70 75 80 Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Met Gln Ala Leu 85 90 95 Gln Thr Leu Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100 105 110 19 121 PRT Homo sapiens PEPTIDE (1)..(121) Clone 2a4 VH 19 Ala Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Lys Pro Gly 1 5 10 15 Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn 20 25 30 Ala Trp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp 35 40 45 Val Gly Arg Ile Lys Ser Lys Thr Asp Gly Gly Thr Ile Asp Tyr Ala 50 55 60 Ala Pro Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Asn 65 70 75 80 Thr Leu Tyr Leu Gln Met Asn Ser Leu Lys Thr Glu Asp Thr Ala Val 85 90 95 Tyr Tyr Cys Thr Thr Trp Asp Gly Asp His Ala Val Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 20 106 PRT Homo sapiens PEPTIDE (1)..(106) Clone 2a4 VK 20 Ala Glu Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Ile Gly Asp 1 5 10 15 Arg Val Ala Ile Ser Cys Gln Ala Ser Gln Asp Ile Gly Asn Tyr Leu 20 25 30 Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Arg Leu Leu Ile Tyr 35 40 45 Asp Ala Ser Asn Leu Glu Ala Gly Val Pro Ser Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Thr Leu Gln Pro Glu 65 70 75 80 Asp Phe Ala Thr Phe Tyr Cys Gln Gln Thr Asp Ser Thr Pro Tyr Thr 85 90 95 Phe Gly Gln Gly Thr Lys Leu Glu Ile Arg 100 105 21 120 PRT Homo sapiens PEPTIDE (1)..(120) Clone 2a5 VH 21 Ala Glu Val Gln Leu Leu Glu Ser Gly Pro Gly Leu Val Lys Pro Ser 1 5 10 15 Glu Thr Leu Ser Leu Thr Cys Thr Val Ser Thr Gly Ser Ile Ser Gly 20 25 30 Tyr Phe Trp Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp 35 40 45 Ile Ala Tyr Ile His Asn Ser Gly Asn Thr Asn Tyr Asn Pro Ser Leu 50 55 60 Arg Ser Arg Val Thr Val Ser Ile Asp Thr Ser Lys Asn Gln Phe Ser 65 70 75 80 Leu Lys Leu Thr Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Asp Gly Gly Gly Trp Asp Thr Tyr Leu Asp Ser Trp Gly Gln 100 105 110 Gly Phe Leu Val Thr Val Ser Ser 115 120 22 105 PRT Homo sapiens PEPTIDE (1)..(105) Clone 2a5 VK 22 Ala Glu Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp 1 5 10 15 Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Thr Val Ser Ser Tyr Leu 20 25 30 Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Val Leu Ile Tyr 35 40 45 Gly Ile Ser Thr Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly Ser 50 55 60 Gly Ser Glu Thr Asp Phe Thr Leu Thr Ile Ser Asn Leu Gln Pro Glu 65 70 75 80 Asp Phe Ala Ile Tyr Tyr Cys Gln Gln Ser Tyr Ser Ser Arg Thr Phe 85 90 95 Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 23 120 PRT Homo sapiens PEPTIDE (1)..(120) Clone 2a13 VH 23 Ala Glu Val Gln Leu Leu Glu Ser Gly Pro Gly Leu Val Lys Pro Ser 1 5 10 15 Glu Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Ser Asp 20 25 30 Tyr Tyr Trp Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp 35 40 45 Ile Gly His Thr Tyr Asp Asn Gly Gly Thr Lys Tyr Asn Pro Ser Leu 50 55 60 Lys Ser Arg Ala Ser Ile Ser Val Asp Thr Ser Lys Asn Gln Val Ser 65 70 75 80 Leu Arg Leu Thr Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Thr Ala Phe Leu Asp Asn Ser Gly Trp Tyr Thr Phe Asp Ser Trp 100 105 110 Gly Gln Gly Ser Leu Val Thr Val 115 120 24 109 PRT Homo sapiens PEPTIDE (1)..(109) Clone 2a13 VK 24 Ala Glu Leu Thr Gln Ser Pro Ser Ser Val Ser Ala Ser Val Arg Asp 1 5 10 15 Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Thr Trp Leu 20 25 30 Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr 35 40 45 Ala Ala Ser Thr Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Gly Ser Leu Gln Pro Glu 65 70 75 80 Asp Val Ala Thr Tyr Tyr Cys Gln Gln Ala Asn Ser Phe Pro Pro Gly 85 90 95 Ala Leu Thr Phe Gly Gly Gly Thr Lys Val Glu Val Lys 100 105 25 121 PRT Homo sapiens PEPTIDE (1)..(121) Clone 2a14 VH 25 Ala Glu Val Gln Leu Leu Glu Gln Ser Gly Gly Gly Val Val Gln Pro 1 5 10 15 Gly Arg Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser 20 25 30 Thr Tyr Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu 35 40 45 Trp Val Ala Val Ile Ser His Asp Gly Ser Asn Lys Phe Tyr Ala Asp 50 55 60 Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Ser Ser Lys Asn Thr 65 70 75 80 Leu Tyr Leu Gln Met Asn Ser Leu Arg Pro Gly Asp Thr Ala Val Tyr 85 90 95 Tyr Cys Ser Arg Trp Asp Arg Arg Ala Glu Tyr Phe Gln Asp Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 26 106 PRT Homo sapiens PEPTIDE (1)..(106) Clone 2a14 VK 26 Ala Glu Leu Thr Gln Ser Pro Ala Phe Met Ser Ala Thr Pro Gly Asp 1 5 10 15 Lys Val Asn Ile Ser Cys Lys Ala Ser Gln Asp Ile Asp Asp Asp Met 20 25 30 Asn Trp Tyr Gln Gln Lys Pro Gly Glu Ala Ala Ile Phe Ile Ile Gln 35 40 45 Glu Ala Thr Thr Leu Val Pro Gly Ile Pro Pro Arg Phe Ser Gly Ser 50 55 60 Gly Tyr Gly Thr Asp Phe Thr Leu Thr Ile Asn Asn Ile Glu Ser Glu 65 70 75 80 Asp Ala Ala Tyr Tyr Phe Cys Leu Gln His Asp Asn Phe Pro Leu Thr 85 90 95 Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 27 118 PRT Homo sapiens PEPTIDE (1)..(118) Clone 2a23 VH 27 Ala Glu Val Gln Leu Leu Glu Ser Gly Ala Gly Val Val Gln Pro Gly 1 5 10 15 Lys Ser Leu Thr Leu Ser Cys Val Gly Ser Gly Phe Thr Phe Ser Ile 20 25 30 Tyr Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp 35 40 45 Val Ala Val Leu Ser Ser Asp Gly Ser Asn Asp Tyr Tyr Ala Asp Ser 50 55 60 Val Lys Gly Arg Phe Ser Ile Phe Arg Asp Thr Ser Lys Asn Ser Leu 65 70 75 80 Asn Leu Leu Met Asn Asn Val Arg Gly Glu Asp Thr Ala Val Tyr Tyr 85 90 95 Cys Ala Arg Asp Gly Asp Gly Ser Phe Phe Tyr Trp Gly Gln Gly Thr 100 105 110 Leu Val Thr Val Ser Ser 115 28 106 PRT Homo sapiens PEPTIDE (1)..(106) Clone 2a23 VK 28 Ala Glu Leu Thr Gln Ser Pro Ala Phe Met Ser Ala Thr Pro Gly Asp 1 5 10 15 Lys Val Asn Ile Ser Cys Lys Ala Ser Gln Asp Ile Asp Asp Asp Met 20 25 30 Asn Trp Tyr Gln Gln Lys Pro Gly Glu Ala Ala Ile Phe Ile Ile Gln 35 40 45 Glu Ala Thr Thr Leu Val Pro Gly Ile Pro Pro Arg Phe Ser Gly Ser 50 55 60 Gly Tyr Gly Thr Asp Phe Thr Leu Thr Ile Asn Asn Ile Glu Ser Glu 65 70 75 80 Asp Ala Ala Tyr Tyr Phe Cys Leu Gln His Asp Asn Phe Pro Leu Thr 85 90 95 Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 29 118 PRT Homo sapiens PEPTIDE (1)..(118) Clone 2a25 VH 29 Ala Glu Val Gln Leu Leu Glu Gln Ser Gly Ala Glu Val Lys Lys Pro 1 5 10 15 Gly Ala Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Met Phe Thr 20 25 30 Cys Tyr Tyr Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu 35 40 45 Trp Met Gly Met Val Asn Pro Thr Gly Gly Ser Ser Ser Tyr Ala Gln 50 55 60 Lys Phe Gln Gly Arg Val Thr Met Thr Lys Gly His Val Thr Ser Thr 65 70 75 80 Val Tyr Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr 85 90 95 Tyr Cys Ala Thr Gly Met Val Arg Gly Asp Glu Trp Gly Gln Gly Thr 100 105 110 Leu Val Thr Val Ser Ser 115 30 106 PRT Homo sapiens PEPTIDE (1)..(106) Clone 2a25 VK 30 Ala Glu Leu Thr Gln Ser Pro Ser Phe Ser Ser Ala Ser Val Gly Asp 1 5 10 15 Arg Val Thr Ile Thr Cys Arg Ala Ser His Gly Ile Ser Ser His Leu 20 25 30 Ala Trp Phe Gln His Lys Pro Gly Lys Ala Pro Asn Leu Leu Ile Tyr 35 40 45 Ala Ala Ser Thr Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly Ser 50 55 60 Gln Ser Gly Thr Glu Phe Thr Leu Thr Ile Thr Ser Leu His Pro Glu 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Leu Tyr Thr Trp Pro Met Gly 85 90 95 Phe Gly Gln Gly Thr Arg Leu Glu Ile Thr 100 105 31 126 PRT Homo sapiens PEPTIDE (1)..(126) Clone 2a30 VH 31 Ala Glu Val Gln Leu Leu Glu Gln Ser Gly Gly Gly Leu Val Gln Pro 1 5 10 15 Gly Gly Ser Leu Arg Leu Ser Cys Thr Ala Ser Gly Phe Thr Leu Thr 20 25 30 Asp Tyr Ser Met Asp Trp Val Arg Gln Ala Pro Gly Lys Gly Pro Glu 35 40 45 Trp Val Gly Arg Ser Arg Asn Lys Ala Asn Ile Tyr Thr Thr Glu Tyr 50 55 60 Ala Ala Ser Val Lys Gly Arg Phe Val Ile Ser Arg Asp Asp Ser Glu 65 70 75 80 Asn Ser Val Tyr Leu Gln Met Asn Asn Val Lys Met Asp Asp Thr Ala 85 90 95 Val Tyr Tyr Cys Ala Arg Gly Glu Gly Ile Phe Tyr Gly Ser Gly Ser 100 105 110 Leu Asp Leu Trp Gly Gln Gly Ala Val Val Thr Val Ser Ser 115 120 125 32 106 PRT Homo sapiens PEPTIDE (1)..(106) Clone 2a30 VK 32 Ala Glu Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp 1 5 10 15 Arg Val Thr Ile Thr Cys Arg Ser Ser Gln Ser Ile Ser Asn Ser Leu 20 25 30 His Trp Phe Gln Gln Glu Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr 35 40 45 Ala Ala Ser Thr Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Asp Phe Thr Leu Ile Ile Thr Gly Leu Gln Pro Glu 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Pro Arg Thr Pro Leu Thr 85 90 95 Phe Gly Gly Gly Thr Lys Val Glu Ile Arg 100 105 33 126 PRT Homo sapiens PEPTIDE (1)..(126) Clone 2a32 VH 33 Ala Glu Val Gln Leu Leu Glu Gln Ser Gly Gly Asp Leu Val Lys Pro 1 5 10 15 Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Thr 20 25 30 Asp Tyr Tyr Met Asn Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu Glu 35 40 45 Trp Ile Ala Tyr Ile Ser Ile Gly Ser Asp Asp Thr Lys Tyr Ala Ala 50 55 60 Ser Val Lys Gly Arg Phe Ser Ile Ser Arg Asp Asn Ala Lys Asn Ser 65 70 75 80 Leu Tyr Leu Gln Met Asn Ser Leu Arg Val Glu Asp Thr Ala Val Tyr 85 90 95 Tyr Cys Gly Arg Gly Gly Gly Tyr Cys Ser Gly Gly Asn Cys Tyr Ser 100 105 110 Ser Asp Tyr Trp Gly Gln Gly Ala Leu Val Thr Val Ser Ser 115 120 125 34 106 PRT Homo sapiens PEPTIDE (1)..(106) Clone 2a32 VK 34 Ala Glu Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Phe Thr Gly Asp 1 5 10 15 Arg Val Ser Ile Thr Cys Arg Ala Ser Gln Gly Ile Gly Asn Ser Leu 20 25 30 Ala Trp Tyr Gln Gln Lys Pro Gly His Val Pro Lys Val Leu Ile Tyr 35 40 45 Asp Ala Ser Thr Leu Ser Ser Gly Ala Pro Ser Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Gly Leu Gln Pro Glu 65 70 75 80 Asp Val Ala Thr Tyr Tyr Cys Gln Lys Tyr Lys Ser Ala Pro Leu Thr 85 90 95 Phe Gly Gly Gly Thr Lys Val Glu Ile Arg 100 105 35 131 PRT Homo sapiens PEPTIDE (1)..(131) Clone 2a33 VH 35 Ala Glu Val Gln Leu Leu Glu Gln Ser Gly Gly Gly Leu Val Gln Pro 1 5 10 15 Gly Arg Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asp 20 25 30 Asp Tyr Ala Met His Trp Val Arg Gln Thr Pro Gly Lys Gly Leu Glu 35 40 45 Trp Val Ser Gly Ile Gly Trp Asn Ser Gly Thr Ile Glu Tyr Ala Asp 50 55 60 Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser 65 70 75 80 Leu Tyr Leu Gln Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Leu Tyr 85 90 95 Tyr Cys Ala Lys Asp Leu His Ser Phe Gly Tyr Cys Ser Gly Arg Ser 100 105 110 Cys Tyr Phe His Pro Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr 115 120 125 Val Ser Ser 130 36 112 PRT Homo sapiens PEPTIDE (1)..(112) Clone 2a33 VK 36 Ala Glu Leu Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly Glu 1 5 10 15 Arg Ala Thr Ile Asn Cys Lys Ser Ser Gln Ser Val Leu Ser Ser Ser 20 25 30 Asn Asn Lys Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Arg Gln Pro 35 40 45 Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val Pro 50 55 60 Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile 65 70 75 80 Ser Ser Leu Gln Ala Glu Asp Val Ala Val Tyr Tyr Cys Gln Gln Phe 85 90 95 Tyr Ser Thr Pro Pro Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Arg 100 105 110 37 118 PRT Homo sapiens PEPTIDE (1)..(118) Clone 2a37 VH 37 Ala Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly 1 5 10 15 Arg Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn 20 25 30 Ser Asp Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp 35 40 45 Val Thr Val Ser Ser Arg Asp Gly Tyr Asp Asn Tyr Tyr Ala Asp Ser 50 55 60 Val Arg Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu 65 70 75 80 Tyr Leu Gln Met Asn Thr Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr 85 90 95 Cys Ala Lys Arg Arg Gly Tyr Ala Phe Asp Ile Trp Gly Gln Gly Thr 100 105 110 Met Val Thr Val Ser Ser 115 38 106 PRT Homo sapiens PEPTIDE (1)..(106) Clone 2a37 VK 38 Ala Glu Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp 1 5 10 15 Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Asn Thr Tyr Leu 20 25 30 Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Glu Leu Leu Ile Tyr 35 40 45 Gly Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Thr Ser Leu Gln Ala Glu 65 70 75 80 Asp Phe Ala Ser Tyr Phe Cys Gln Gln Ser Asp Ser Thr Pro Arg Thr 85 90 95 Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 39 128 PRT Homo sapiens PEPTIDE (1)..(128) Clone 2a40 VH 39 Ala Glu Val Gln Leu Leu Glu Gln Ser Gly Gly Gly Leu Val Lys Pro 1 5 10 15 Gly Gly Ser Leu Arg Leu Ser Cys Thr Ala Ser Gly Phe Thr Phe Ser 20 25 30 Gly Phe Thr Phe Ser Asp His Tyr Met Ser Trp Ile Arg Gln Ala Pro 35 40 45 Gly Lys Gly Leu Glu Leu Val Ser Tyr Ile Ile Ser Asn Gly Tyr Thr 50 55 60 Asn Tyr Pro Asp Ser Val Lys Gly Arg Phe Thr Val Ser Arg Asp Asn 65 70 75 80 Ala Arg Lys Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg Val Glu Asp 85 90 95 Thr Ala Ile Tyr Tyr Cys Ala Arg Gly Leu Ser Pro Ser Ile Ala Gly 100 105 110 Asp Gly Phe Asp Ile Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser 115 120 125 40 106 PRT Homo sapiens PEPTIDE (1)..(106) Clone 2a40 VK 40 Ala Glu Leu Thr Gln Ser Pro Ala Phe Met Ser Ala Thr Pro Gly Asp 1 5 10 15 Lys Val Asn Ile Ser Cys Lys Ala Ser Gln Asp Ile Asp Asp Asp Met 20 25 30 Asn Trp Tyr Gln Gln Lys Pro Gly Glu Ala Ala Ile Phe Ile Ile Gln 35 40 45 Glu Ala Thr Thr Leu Val Pro Gly Ile Pro Pro Arg Phe Ser Gly Ser 50 55 60 Gly Tyr Gly Thr Asp Phe Thr Leu Thr Ile Asn Asn Ile Glu Ser Glu 65 70 75 80 Asp Ala Ala Tyr Tyr Phe Cys Leu Gln His Asp Asn Phe Pro Leu Thr 85 90 95 Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 41 116 PRT Homo sapiens PEPTIDE (1)..(116) Clone 2b1 VH 41 Ala Glu Val Gln Leu Leu Glu Ser Gly Pro Arg Leu Val Lys Pro Ser 1 5 10 15 Gln Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Asp Ser Leu Asn Asn 20 25 30 Ala Ser His Tyr Trp Ala Trp Ile Arg Gln Pro Ala Gly Lys Gly Leu 35 40 45 Glu Trp Ile Gly Arg Ile His Arg Gly Gly Ser Thr Asn Tyr Asn Pro 50 55 60 Ser Leu Gln Ser Arg Val Thr Ile Ser Met Asp Glu Ser Lys Asn Gln 65 70 75 80 Phe Ser Leu Arg Leu Asn Ser Val Thr Ala Ala Asp Thr Ala Val Tyr 85 90 95 Tyr Cys Ala Arg Asp Pro Pro Lys Ala Trp Gly Pro Gly Ile Leu Val 100 105 110 Thr Val Ser Ser 115 42 105 PRT Homo sapiens PEPTIDE (1)..(105) Clone 2b1 VK 42 Ala Glu Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp 1 5 10 15 Arg Val Thr Ile Ala Cys Arg Ala Ser Gln Ser Ile Ser Asn His Leu 20 25 30 Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Val Leu Ile Tyr 35 40 45 Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu 65 70 75 80 Asp Phe Ala Ile Tyr Tyr Cys Gln Gln Ser Tyr Ser Asn Ala Asp Phe 85 90 95 Gly Pro Gly Thr Lys Val Asp Ile Lys 100 105 43 121 PRT Homo sapiens PEPTIDE (1)..(121) Clone 2b3 VH 43 Ala Glu Val Gln Leu Leu Glu Ser Gly Ala Glu Val Lys Lys Pro Gly 1 5 10 15 Ala Ser Val Thr Val Ser Cys Lys Ala Ser Gly Tyr Ser Phe Ser Asp 20 25 30 Tyr Tyr Ile His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp 35 40 45 Met Gly Ile Ile Asn Pro Ser Ala Gly Thr Thr Thr Tyr Lys Gln Lys 50 55 60 Phe Gln His Arg Val Thr Leu Thr Arg Asp Thr Ser Thr Asn Thr Ala 65 70 75 80 Tyr Met Lys Leu Tyr Asn Leu Thr Pro Asp Asp Thr Ala Ile Phe Phe 85 90 95 Cys Ala Arg Gly Ser Gly Gly Ser Arg Ser Glu Tyr Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 44 106 PRT Homo sapiens PEPTIDE (1)..(106) Clone 2b3 VK 44 Ala Glu Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp 1 5 10 15 Gly Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Thr Tyr Leu 20 25 30 Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Met Tyr 35 40 45 Ala Ala Ser Thr Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu 65 70 75 80 Asp Phe Ala Ala Tyr Tyr Cys Gln Gln Ala Arg Ser Phe Pro Tyr Thr 85 90 95 Phe Gly Gln Gly Thr Arg Leu Glu Ile Arg 100 105 45 121 PRT Homo sapiens PEPTIDE (1)..(121) Clone 2b4 VH 45 Ala Glu Val Gln Leu Leu Glu Ser Gly Gly Glu Val Lys Lys Pro Gly 1 5 10 15 Ser Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Pro Phe Ser Ser 20 25 30 Tyr Ala Ile His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp 35 40 45 Met Gly Ala Ile Ile Pro Phe Leu Gly Arg Ala Lys Tyr Ala Gln Lys 50 55 60 Phe Glu Gly Arg Val Thr Ile Thr Ala Asp Gly Ser Met Ser Thr Ala 65 70 75 80 Tyr Met Glu Val Ser Ser Leu Arg Ser Asp Asp Thr Ala Met Tyr Tyr 85 90 95 Cys Ala Arg Asp Arg Gly Glu Leu Leu Leu Arg Met Asp Val Trp Gly 100 105 110 Gln Gly Thr Ala Val Thr Val Ser Ser 115 120 46 106 PRT Homo sapiens PEPTIDE (1)..(106) Clone 2b4 VK 46 Ala Glu Leu Thr Gln Ser Pro Ala Phe Met Ser Ala Thr Pro Gly Asp 1 5 10 15 Lys Val Asn Ile Ser Cys Lys Ala Ser Gln Asp Ile Asp Asp Asp Met 20 25 30 Asn Trp Tyr Gln Gln Lys Pro Gly Glu Ala Ala Ile Phe Ile Ile Gln 35 40 45 Glu Ala Thr Thr Leu Val Pro Gly Ile Pro Pro Arg Phe Ser Gly Ser 50 55 60 Gly Tyr Gly Thr Asp Phe Thr Leu Thr Ile Asn Asn Ile Glu Ser Glu 65 70 75 80 Asp Ala Ala Tyr Tyr Phe Cys Leu Gln His Asp Asn Phe Pro Leu Thr 85 90 95 Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 47 128 PRT Homo sapiens PEPTIDE (1)..(128) Clone 2b5 VH 47 Ala Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Val Val Gln Pro Gly 1 5 10 15 Arg Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Leu Ser Thr 20 25 30 Tyr Ala Met His Trp Val Arg Gln Ala Pro Gly Arg Gly Leu Glu Trp 35 40 45 Val Ala Val Ile Ser Tyr Asp Gly Asp His Lys Phe Tyr Ala Asp Ser 50 55 60 Met Lys Gly Arg Phe Ala Ile Ser Arg Asp Thr Ser Thr Asn Thr Leu 65 70 75 80 Tyr Leu Glu Val Asn Ser Leu Lys Ile Glu Asp Thr Ala Val Tyr Tyr 85 90 95 Cys Ala Arg Asp Arg Asp Arg Arg Gly Gly Tyr Val Phe Ser Thr Thr 100 105 110 Gly Gly Leu Asp Ser Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 125 48 111 PRT Homo sapiens PEPTIDE (1)..(111) Clone 2b5 VK 48 Ala Glu Leu Thr Gln Ser Pro Leu Ser Leu Ala Val Thr Pro Gly Glu 1 5 10 15 Pro Ala Ser Ile Ser Cys Arg Thr Ser Gln Ser Leu Leu His Ser Asn 20 25 30 Gly Tyr Thr Tyr Leu Asp Trp Tyr Leu Gln Lys Pro Gly Gln Ser Pro 35 40 45 Gln Leu Leu Ile Tyr Leu Ala Ser Asn Arg Ala Ser Gly Val Pro Asp 50 55 60 Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Glu Ile Ser 65 70 75 80 Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Met Gln Pro Leu 85 90 95 Gln Thr Pro Val Thr Phe Gly Gln Gly Thr Lys Val Glu Val Lys 100 105 110 49 123 PRT Homo sapiens PEPTIDE (1)..(123) Clone 2b7 VH 49 Ala Glu Val Gln Leu Leu Glu Gln Ser Gly Ala Glu Val Lys Lys Pro 1 5 10 15 Gly Glu Ser Leu Lys Ile Ser Cys Arg Ala Ser Gly Tyr Ser Phe Ser 20 25 30 Leu Phe Trp Val Ala Trp Val Arg Gln Met Pro Gly Gln Gly Leu Glu 35 40 45 Trp Met Ala Ile Ile Tyr Pro Gly Asp Ser Asp Thr Thr Tyr Ser Pro 50 55 60 Ser Phe Glu Gly Gln Val Asn Val Ser Val Asp Lys Pro Ile Ser Thr 65 70 75 80 Ala Tyr Leu Gln Trp Ser Ser Leu Lys Ala Ser Asp Thr Ala Met Tyr 85 90 95 Tyr Cys Ala Arg Arg Arg Ser Ser Asp Arg Arg Asp Ala Phe Asp Ile 100 105 110 Trp Gly Pro Gly Thr Met Val Thr Val Ser Ser 115 120 50 106 PRT Homo sapiens PEPTIDE (1)..(106) Clone 2b7 VK 50 Ala Glu Leu Thr Gln Ser Pro Ala Phe Met Ser Ala Thr Pro Gly Asp 1 5 10 15 Lys Val Asn Ile Ser Cys Lys Ala Ser Gln Asp Ile Asp Asp Asp Met 20 25 30 Asn Trp Tyr Gln Gln Lys Pro Gly Glu Ala Ala Ile Phe Ile Ile Gln 35 40 45 Glu Ala Thr Thr Leu Val Pro Gly Ile Pro Pro Arg Phe Ser Gly Ser 50 55 60 Gly Tyr Gly Thr Asp Phe Thr Leu Thr Ile Asn Asn Ile Glu Ser Glu 65 70 75 80 Asp Ala Ala Tyr Tyr Phe Cys Leu Gln His Asp Asn Phe Pro Leu Thr 85 90 95 Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 51 128 PRT Homo sapiens PEPTIDE (1)..(128) Clone 2b9 VH 51 Ala Glu Val Gln Leu Leu Glu Ser Gly Pro Gly Leu Val Gln Pro Ser 1 5 10 15 Gln Thr Leu Ser Leu Thr Tyr Ala Ile Ser Gly Asp Ser Val Ser Ser 20 25 30 Asn Ser Ala Ala Trp Thr Trp Ile Arg Gln Ser Pro Ser Arg Gly Leu 35 40 45 Glu Trp Leu Gly Met Thr Tyr Tyr Arg Ser Gln Trp Tyr His Glu Tyr 50 55 60 Ala Val Ser Leu Lys Ser Arg Ile Thr Ile Asn Ala Asp Thr Ser Asn 65 70 75 80 Asn Gln Phe Ser Leu Gln Val Asn Ser Val Thr Pro Glu Asp Thr Ala 85 90 95 Leu Tyr Tyr Cys Ala Arg Ala Arg Phe Val Gly Asp Thr Thr Gly Tyr 100 105 110 Tyr Thr Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 125 52 107 PRT Homo sapiens PEPTIDE (1)..(107) Clone 2b9 VK 52 Ala Glu Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly Glu 1 5 10 15 Arg Gly Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Ser Tyr 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Ala Gly Gln Ala Pro Arg Leu Leu Ile 35 40 45 Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu Pro 65 70 75 80 Glu Asp Phe Ala Val Tyr Tyr Cys Gln Arg Tyr Gly Thr Ser Pro Lys 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Phe Lys 100 105 53 127 PRT Homo sapiens PEPTIDE (1)..(127) Clone 2b10 VH 53 Ala Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Val Val Gln Pro Gly 1 5 10 15 Arg Ser Leu Lys Leu Ser Cys Thr Ala Ser Thr Phe Thr Phe Thr Asn 20 25 30 Tyr Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp 35 40 45 Val Ala Leu Ile Ser Asn Asp Gly Ser Lys Thr Tyr Tyr Thr Asp Ser 50 55 60 Val Arg Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu 65 70 75 80 Phe Leu Gln Met Asn Ser Leu Arg Thr Glu Asp Thr Ala Val Tyr His 85 90 95 Cys Ala Arg Val Lys Leu Gln Gly Ser Phe Asn Val Tyr Tyr Tyr Tyr 100 105 110 Gly Leu Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser 115 120 125 54 110 PRT Homo sapiens PEPTIDE (1)..(110) Clone 2b10 VK 54 Ala Glu Leu Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly Glu 1 5 10 15 Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Leu His Ser Asp 20 25 30 Gly Tyr Asn Tyr Phe Asp Trp Tyr Leu Gln Lys Pro Gly Gln Ser Pro 35 40 45 Gln Leu Leu Ile Tyr Leu Gly Ser Asn Arg Ala Ser Gly Val Pro Asp 50 55 60 Arg Phe Ser Gly Ser Gly Ser Asp Thr Asp Phe Thr Leu Lys Ile Ser 65 70 75 80 Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Met Gln Thr Leu 85 90 95 Gln Thr Leu Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 110 55 118 PRT Homo sapiens PEPTIDE (1)..(118) Clone 2b17 VH 55 Ala Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly 1 5 10 15 Arg Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn 20 25 30 Ser Asp Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp 35 40 45 Val Thr Val Ser Ser Arg Asp Gly Tyr Asp Asn Tyr Tyr Ala Asp Ser 50 55 60 Val Arg Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu 65 70 75 80 Tyr Leu Gln Met Asn Thr Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr 85 90 95 Cys Ala Lys Arg Arg Gly Tyr Ala Phe Asp Ile Trp Gly Gln Gly Thr 100 105 110 Met Val Thr Val Ser Ser 115 56 106 PRT Homo sapiens PEPTIDE (1)..(106) Clone 2b17 VK 56 Ala Glu Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp 1 5 10 15 Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Asn Thr Tyr Leu 20 25 30 Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Glu Leu Leu Ile Tyr 35 40 45 Gly Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Thr Ser Leu Gln Ala Glu 65 70 75 80 Asp Phe Ala Ser Tyr Phe Cys Gln Gln Ser Asp Ser Thr Pro Arg Thr 85 90 95 Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 

1. A human monoclonal antibody or antigen binding fragments thereof that exhibits immunological binding affinity for a hepatitis C virus (HCV) E1 antigen, wherein said monoclonal antibody comprises an amino acid sequence homologous to the binding portion of a human antibody Fab molecule obtained from a combinatorial antibody library.
 2. A monoclonal antibody according to claim 1, wherein the monoclonal antibody reacts with complexed HCV E1/E2 antigen.
 3. A monoclonal antibody according to claim 1, comprising any of the amino acid sequences in the enclosed Sequence listing.
 4. The monoclonal antibody according to claim 1, fused with a further substance for diagnostic or therapeutic purposes.
 5. An immunological reagent comprising the monoclonal antibody according to claim
 1. 6. An immunological assay comprising the reagent according to claim
 5. 7. A vaccine composition against HCV infection, comprising the monoclonal antibody according to claim 1 and a pharmaceutically acceptable vehicle for passive immunization.
 8. A vaccine composition according to claim 7, further comprising antibodies, or antigen binding fragments, against HCV E2 antigen.
 9. A vaccine composition according to claim 7, further comprising antibodies, or antigen binding fragments, against the hypervariable region (HVR) of HCV E2.
 10. A method of treating a subject against HCV infection in a therapeutic or prophylactic purpose, comprising administration of the vaccine composition according to claim 7 to subjects in need thereof.
 11. A monoclonal antibody according to claim 2, comprising any of the amino acid sequences listed in the enclosed sequence listing.
 12. A monoclonal antibody according to claim 2 fused with a further substance for diagnostic or therapeutic purposes.
 13. A monoclonal antibody according to claim 3, fused with a further substance for diagnostic or therapeutic purposes
 14. An immunological reagent comprising the monoclonal antibody according to claim
 3. 15. A vaccine composition against HCV infection, comprising the monoclonal antibody according to claim 3, and a pharmaceutically acceptable vehicle for passive immunization.
 16. A method of treating a subject against HCV infection in a therapeutic or prophylactic purpose, comprising administration of the vaccine composition according to claim 8 to subjects in need thereof.
 17. A method of treating a subject against HCV infection in a therapeutic or prophylactic purpose, comprising administration of the vaccine composition according to claim 9 to subjects in need thereof. 